细叶百合鳞茎在低温解除休眠过程中茎尖细胞超微结构的变化

 研究了细叶百合鳞茎低温处理打破休眠过程中的形态变化及茎尖细胞超微结构的变化。结果表明,低温处理0 ~ 12 d,鳞茎物质和能量代谢较弱,鳞茎不能萌发;12 ~ 48 d,鳞茎萌发率较低,线粒体及高尔基体分泌囊泡增多,细胞通过质膜内吞及胞间连丝进行物质交流;48 ~ 72 d,核仁松散,内质网有序排列,在液泡膜与质膜边缘形成内质网桥,内质网桥一直延伸到核膜方向,胞内及胞间联系已经建立;出芽时,内质网桥延伸到胞间连丝,细胞器内部结构及数量增加,休眠完全打破。     

苹果梨花芽孕育期生长锥细胞超微结构的观察

利用透射电镜对苹果梨花芽孕育期芽内生长锥细胞进行了观察，结果显示，在花芽孕育形成的过程中，生长锥细胞的超微结构发生了一系列变化：蛋白体和脂类小滴逐渐增加；核外膜向外延伸到胞质中与内质网相连，其上分布的核糖体增多；线粒体和内质网数目丰富且分布均匀；核糖体大量游散在胞质中；质体呈浅裂，中央缢缩；微管分布于靠近质膜处等。以上超微结构的变化表现出细胞旺盛分裂和分化的特点，反映了花芽孕育过程中的细胞学特征。     

蛹虫草继代中菌株退化早期外部识别标志研究

以蛹虫草菌株W5继代培养(8 d转接1代)1～5代菌株(W5-1～W5-5)为研究对象，观察蛹虫草继代培养过程中菌落形态、细胞超微结构和活性氧含量的变化，以期揭示继代培养过程中蛹虫草菌株退化的早期外部识别标志。结果表明：蛹虫草W5菌株退化是个渐变的过程，继代培养至3代菌落形态、菌丝和孢子形态开始出现变化，节律环和颜色变化以及菌丝表面的缢裂和凹陷是3代的显著外部特征，可作为蛹虫草菌株退化的早期菌落形态识别标志。在蛹虫草W5菌株继代培养过程中，菌丝的细胞壁逐渐变薄，细胞核逐渐变大，线粒体逐渐肿胀变形，液泡逐渐增大，膜边体逐渐减少，圆球体逐渐溶解，形成脂滴和嗜锇颗粒。可以通过细胞中核仁是否显现、有无膜边体、有无液泡融合和自噬现象来早期判断菌株的退化程度。从3代开始菌丝细胞NBT染色颜色逐渐加深，反映细胞活性氧含量增加。细胞H₂O₂含量测定表明，3代H₂O₂含量显著高于前2代。基于线粒体与活性氧的紧密关系，认为线粒体结构和数量上的改变可能是蛹虫草退化的重要原因之一。     

平菇孢子萌发的细胞学观察

在进行食用菌杂交育种时,亲本的孢子萌发必须和菌丝发育阶段相近,才能通过菌丝融合形成双核菌丝,产生新类型。为了使亲本孢子萌发和菌丝发育阶段相近,研究孢子萌发到双核菌丝形成的细胞学过程是十分重要的。我们采用凹玻片悬滴培养平菇孢子,成功地进行了从孢子萌发到双核菌丝形成的显微观察。从实践中体会到,用此法培养平菇孢子,比采用平皿、试管斜面、液体浅层培养法更为简便和省料,且适于在光镜下直接进行活体观察。     

冰核细菌对仁用杏花粉超微结构的影响

 应用透射电镜对几个仁用杏品种接种冰核细菌并低温处理后的花粉超微结构进行观察, 发现冰核细菌对花粉的超微结构有很大影响: ( 1) 使花粉壁的覆盖层在某些区域解体消失或整个花粉外壁缺失; ( 2) 使花粉中的细胞器受到破坏, 包括线粒体被膜不完整, 嵴呈囊泡状; 内质网呈同心圆卷绕或折叠状排列包围部分细胞质而形成膜内含物; 核糖体数量明显减少等。因为花粉中细胞器的破坏, 花粉细胞内部形成一些空腔, 花粉失去生活力。接种冰核细菌并低温处理比单纯低温处理对花粉超微结构的破坏程度加重。     

平菇呼吸强度初探

有关平菇呼吸强度的研究,迄今尚未见有报道,因此笔者对平菇吸氧量进行了测定,探讨其子实体、菌丝和孢子三个阶段的呼吸强度及温度对子实体吸氧量的影响,为平菇吸氧量提出明确的生理参数,以推算出平菇在不同的各个阶段所需氧的量,提供给有关人员参考。材料和方法菌种:平菇404,引自华中农业大学真菌研究室。孢子:取新鲜子实体弹射的孢子,弹射时间不得超过24小时,以免失去活性的孢子     

灵芝的磁生物效应及其超微结构的研究

本文首次报道了用电镜观察到喷浇磁水的灵芝其菌肉细胞密度比对照组高１／３左右,且厚壁细胞彘薄壁细胞多１／３左右,而对照组则是厚壁细胞比薄膜细胞少１／３左右。在喷浇磁水的灵芝菌管细胞内不可见到较多的线粒体和密集的糖原颗粒。孢子内外结构及大小基本相同。并阐明灵芝超微结构２的变化与磁生物效应的关系。     

西瓜小孢子有丝分裂前后质体和线粒体的变化

为揭示西瓜小孢子有丝分裂前后质体和线粒体的变化以及它们向生殖细胞和营养细胞分配的特点,用透射电子显微技术对西瓜不同发育阶段小孢子及早期二细胞花粉的质体和线粒体进行比较研究。结果表明,小孢子的质体结构简单,不积累淀粉等贮藏物质,为原质体;二细胞花粉的质体不含内部膜系统,但积累大量淀粉,表明小孢子有丝分裂引起原质体向造粉体分化。二细胞花粉的质体都在营养细胞中,生殖细胞不含质体,显然,小孢子分裂过程中质体只分配给营养细胞而不分配给生殖细胞,这一不均等分配方式决定质体呈单亲母系遗传,据此,将西瓜的质体遗传细胞学机制归类为番茄型。小孢子的线粒体内嵴较少,营养细胞和生殖细胞的线粒体内嵴较多,显示小孢子分裂后线粒体结构复杂化。     

蘑菇菌种的长期贮存方法

降低细胞代谢速度是营养饥饿、除氧、液氮保藏及冰冻干燥等方法的理论基础。这些方法单独或混和使用。脱水保藏法水是细胞代谢不可缺少的物质,因此脱水可限制代谢活动。其方法:将真菌菌丝或孢子吸附在土粒、硅肢及纤维材料(滤纸等)上,然后冷冻干燥。据报导,蘑菇孢子可长期保藏在土粒或滤纸中,然而未作贮藏后的各种生理特征试验。我们将香菇、平菇的菌丝片断和孢子吸附在无菌的硅胶上,贮于5℃或-20℃中,一年后它们均存活,但没对其活力及生理特性进行测试。     

保水剂在平菇栽培中的应用研究

为解决平菇栽培过程中培养料失水以及补水问题，提高平菇产量，本试验将不同类型（MP3005KM、MP3005KB、MP—KR、MP—KS）、不同浓度（0．1％、0．5％、1％、1．5％、2％）的保水剂吸水制成凝胶颗粒后进行拌料处理，并对平菇茵丝生长、出菇以及产量等进行观察与测定研究。结果表明，保水剂使用浓度为0．5％时效果最好，可使菌丝布满袋时间提前2d～5d，出菇提前4d-6d，产量提高20％左右。保水剂使用浓度≥1％时，菌丝生长受到抑制、出菇时间延迟，且浓度越大，菌丝生长及出菇速度越缓慢，对产量亦产生不利影响。在保水剂对菌丝生长、出菇以及产量的影响趋势方面，4种保水剂之间并无明显区别。由此可见，选择适宜浓度的保水剂制成凝胶颗粒进行拌料处理，是解决平菇栽培过程中培养料失水以及补水问题，提高平菇产量的有效途径，值得推广应用。     

Morphology and growth capacity of bone marrow stromal cells in vitro from APBSCT patients post pretreatment

AIM: To investigate the morphology and capacity of bone marrow colony forming unity-fibroblast (CFU-F) from APBSCT patients before and after pretreatment. METHODS: 21 case peripheral blood stem cell transplantation (PBSCT) patients were treated with pretreatment. The changes of morphology of the bone marrow stromal cells were assayed by light microscope and electron microscope, respectively. The numbers of CFU-F were assayed by Dexter type. RESULTS: The bone marrow stromal cells occured different types of morphology from PBSCT patients treated with chemotherapy or chemotherapy-TBI pretreatment, respectively, compared with controls. The transmission electron microscope showed that the endoplasmic reticula was dilated, the matrix of mitochondria appeared pale and the cristae of mitochondria became shorter in stromal cells from chemotherapy-TBI patients compared with those of controls. The structure of mitochondria from combined chemotherapy-TBI pretreatment appeared severe degeneration and disorder. The numbers of CFU-F from combined radiation-chemotherapy injury were significantly decreased compared with that before pretreatment and the chemotherapy injury (P<0.01), respectively. CONCLUSION: The change of cell morphology and capacity of CFU-F for bone marrow stromal cells is one of impairment injury mechanism of bone marrow hematopoietic inductive microenvironment from PBSCT patients post pretreatment.     

西瓜成熟花粉的超微结构

为了解西瓜成熟花粉的超微结构特点,特别是营养细胞和生殖细胞之间的关系,选取刚开花的西瓜雄花采集花粉,2.5%戊二醛固定并制成超薄切片,透射电镜观察照相。结果表明,西瓜成熟花粉粒中营养细胞含丰富脂类储藏物质和内质网,内质网堆叠,周围和间隙中充满脂类物质;含丰富高尔基体和大量小泡;质体很少。营养核通常深度开裂,产生多数裂片,裂片伸向生殖细胞。生殖细胞流线型,细胞壁电子透明,细胞核伸长,细胞质含高尔基体、内质网,不含质体。生殖细胞表面产生波状突起,波峰背离营养核。生殖细胞一端伸入营养核裂片间隙,2者构成雄性生殖单位。     

Ultrastructural changes and significance of endometriotic rat model with HCG treatment

AIM: To investigate whether and how human chorionic gonadotropin (HCG) treatment ameliorates endometriosis in the endometriotic rat model. METHODS: The rat model of endometriosis was established and the model rats were divided into 4 groups. The rats in HCG groups were treated with 19.4, 25.8 and 51.6 IU/100 g of HCG every day (low-dose HCG, medium-dose HCG and high-dose HCG, respectively). The rats in control group were treated with 0.9% NaCl. After 15 days (3 estrous cycles), the ectopic lesion volume and ultrastructural characteristics in eutopic and ectopic endometria were investigated. RESULTS: After HCG treatment, the volume of endometriotic lesions was significantly smaller than that before treatment. Numerous and mitochondrial, endoplasmic reticulum and ribosomes were observed in the cytoplasm of eutopic and ectopic endometrium before treatment. After treatment, some cell structures were not clear, and mitochondrial cristae decreased or disappeared partly. Some cells were densed and shrinkage, autophagosome in cytoplasm increased, and mitochondria and endoplasmic reticulum swelt. CONCLUSION: HCG therapy appears to be an effective treatment for endometriosis in rats attributed to its influence on cell metabolism dysfunction of eutopic and ectopic endometria.     

Immunohistochemical localization of phospholipase D in strawberry (Fragaria ananassa Duch.) fruits

Phospholipase D, the key enzyme involved in membrane phospholipid catabolism, was localized in strawberry (Fragaria ananassa Duch.) fruits during development using immunohistochemical techniques. Typical structural features such as the nucleus, cell wall, plasma membrane, mitochondria and vacuole were noticeable in cells of young strawberry fruits. Cytoplasmic structural features were less pronounced in fully grown and turning red fruits, except cell wall, which was considerably enlarged. Also, extensive microvesiculation from the plasma membrane was noticeable in ripening fruits. The number of visible gold particles indicative of phospholipase D increased during development. Phospholipase D was found to be associated with membranous structures and vacuole. Interestingly, gold-particles could also be observed in the cell wall space in ripening fruits suggesting that cytosolic contents may have leaked into the cell wall space as a result of membrane deterioration and loss of compartmentalization. The role of phospholipase D during development and ripening of strawberry, a non-climacteric fruit, is discussed.     

Mitochondria associated endoplasmic reticulum membranes in cisplatin induced ovarian cancer cell apoptosis

AIM: To explore the structural change of mitochondria associated endoplasmic reticulum membranes (MAMs) in SKOV3 cells exposed to cisplatin. METHODS: The SKOV3 cells were treated with cisplatin at concentration of 6 mg/L. The protein levels of active caspase-3, as well as the colocalization of B-cell receptor-associated protein 31 (BAP31) and voltage-dependent anion channel protein 1 (VDAC1) in the SKOV3 cells were determined by the method of indirect immunofluorescence. The apoptotic rate of the SKOV3 cells was analyzed by flow cytometry. The structural change of MAMs was observed under transmission electron microscope. RESULTS: Under the confocal microscope, we found that cisplatin increased the protein levels of active caspase-3 as well as colocalization of BAP31 and VDAC1 in the SKOV3 cells. The results of flow cytometry demonstrated that cisplatin increased the apoptotic rate of the SKOV3 cells (P<0.05). The results of transmission electron microscopy showed that cisplatin induced increase in mitochondrial-associated membrane structures (P<0.05). CONCLUSION: Cisplatin induces SKOV3 cell apoptosis with increased MAMs contacts. MAMs may play a role in cisplatin induced SKOV3 cell apoptosis.     

Effects of artesunate on membrane surface morphology of human gastric cancer SGC-7901 cells observed under atomic force microscope

AIM: To detect the membrane surface morphology of cancer cells treated with artesunate (ART) under atomic force microscope (AFM). METHODS: Human gastric cancer SGC-7901 cells were cultured and treated with different concentrations of ART for 24 h. The membrane surface morphology, three-dimensional structures and ultrastructural changes of SGC-7901 cells were observed under AFM. The apoptosis of SGC-7901 cells was detected by flow cytometry. RESULTS: The AFM images revealed that the cell nuclear area was full and the surface of cell membrane was flat and smooth in control group. Compared with control group, the cell nuclear area collapsed and suffered atrophy, and the membrane surface had more pores in ART treatment groups. More pores were observed and the diameter of the pores was increased with the increase in ART concentration. The cell membrane ultrastructure showed that the particles in control group had an intensive distribution, some were cord-shaped and some gathered into a mass. The particles in ART treatment groups were fewer and distributed sparsely. Moreover, there were obvious lacunae in the surface of cell membrane. Quantitive measurement found that the height of cell nucleus area was decreased, and the surface root mean square roughness (Rq) and the surface average roughness (Ra) became smaller in ART treatment groups compared with control group. The apoptotic rates of the cells in ART treatment groups were increased with the increase in ART concentration, and were all significantly higher than that in control group. CONCLUSION: Our findings emphasize the significance of AFM in exploring the changes of the membrane surface morphology at nano-scale resolution following the treatment with anticancer drugs, which could not only identify the specific characteristics of morphological changes of the tumor cells, but also provide micromorphological references to reveal the mechanisms of anticancer drugs.     

梨果实发育中Ca2+在果肉细胞的定位及变化研究*

 用焦锑酸钾沉淀的细胞化学方法, 研究了􀀂 幸水 梨果实发育中果肉细胞的焦锑酸Ca²⁺ 定位变化及其与细胞超微结构的关系。结果表明: ( 1) 在未受精之前, 果肉细胞内未检测到Ca²⁺ 沉淀颗粒, 细胞核内的染色质少且染色淡, 细胞质的细胞器数量也少; ( 2) 受精后果肉细胞呈现大量的Ca²⁺沉淀颗粒, 主要分布在细胞核、细胞质、质体以及叶绿体外膜上, 含Ca²⁺沉淀颗粒的质体非常膨大, 导管和初期发育的石细胞内也密集分布Ca2+ 沉淀颗粒; ( 3) 受精1 周后果肉细胞的Ca²⁺移向细胞之间的连接处; ( 4) 生理落果的细胞和导管中Ca²⁺没有或极少, 但有的细胞内沿液泡膜有Ca²⁺分布; ( 5) 受精3~ 4 周后, 果肉细胞中很难检测到Ca²⁺沉淀颗粒, 此状态一直持续到果实采收, 但果实腐烂前Ca²⁺沉淀颗粒沿果肉细胞 壁两侧出现。就Ca2+ 在果实发育中的作用及与细胞超微结构的关系等进行了讨论。     

Mechanisms of augmenter of liver regeneration in promoting damaged hepatocyte proliferation

AIM: To investigate the mechanisms of augmenter of liver regeneration (ALR) in promoting damaged hepatocyte proliferation.METHODS: The effects of Kupffer cell condition medium (KCCM+) stimulated by ALR on damaged hepatocyte proliferation were studied by MTT. The localization of ALR binding to Kupffer cell membrane and in intact rat liver was studied by immunohistochemistry. The IL-6 expression in Kupffer cells stimulated with ALR was observed by immunohistochemistry. RESULTS: The proliferation of damaged hepatocytes stimulated with KCCM+ was increased significantly. ALR immunostaining particles in plasm of hepatocyte were found in intact liver. The rough immunostaining particles of ALR were seen on the surface of Kupffer cell membrane. Immunostaining particles of IL-6 in Kupffer cells induced by ALR increased. CONCLUSION: ALR promotes proliferation of damaged hepatocytes indirectly by stimulating Kupffer cells.